Absorption, Distribution, Metabolism and Leaching of [14C] ABA during Culture of Apple Embryos

Barthe, Ph. and Bulard, C. 1987. Absorption, distribution, metabolismand leaching of [¹⁴C] ABA during culture of apple embryos.—J.exp. Bot. 38: 1002–1011. It has been known for some time that dormant embryos, laid flaton damp filter-paper one cotyledon only being in direct contactwith it (C/2M mode of culture), exhibit unequal growth and greeningof their cotyledons. The aim of this work was to investigatewhether this particular mode of culture led to detectable differencesbetween the two cotyledons in the distribution and metabolismof [¹⁴C] abscisic acid (ABA). Two different approaches wereused, the material in both cases being dormant embryos of Pyrusmalus L. cv. Golden Delicious cultured at 23°C in darkness.In a first experiment, the embryos were cultured directly inthe C/2M mode and in the presence of 10^–2 mol m^–3[¹⁴C] ABA. In these conditions marked differences in the distributionof radioactivity between the lower (LC) and upper (UC) cotyledonappear after 24 h of culture. After 5 d, amounts of total radioactivitywere four times higher in LC than in UC, and the level of ABAwas three times higher. Metabolism was extremely active in LC,since certain metabolites were found in relative percentages(percentages with respect to the total radioactivity of thecotyledon) equivalent to (esters, glucosides), or even higher(dihydrophaseic acid, DPA) than those found in UC. It is suggestedthat this high level of metabolism in LC limits the availabilityof the transportable molecule, that is to say ABA. In a secondexperiment double culturing was carried out. The embryos werefirst cultured in the presence of 10^–2 mol m^–3 [¹⁴C]ABA in conditions which ensured equal distribution of ABA andits metabolites between the two cotyledons. After 5 d they weretransferred to an ABA-free medium and cultured in the C/2M mode.After 2 d and 5 d of culture, dissymmetry between the two cotyledonswas again noted; UC this time containing greater amounts ofradioactivity than LC. The differences, however, were less markedthan in the first experiment. This dissymmetry is due to leachingof a certain amount of radioactivity from LC, which is onlypartially compensated for by migration from UC. Key words: ¹⁴C-ABA distribution, leaching and metabolism, embryo culture, embryo dormancy